PMID- 15598882
OWN - NLM
STAT- MEDLINE
DCOM- 20050718
LR  - 20210108
IS  - 0021-924X (Print)
IS  - 0021-924X (Linking)
VI  - 136
IP  - 3
DP  - 2004 Sep
TI  - Identification and characterization of the hypoxia-responsive element of the 
      human placental 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase gene.
PG  - 273-7
AB  - The placenta-type isozyme of human 
      6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase (HP2K, identical to PFKFB3) 
      is expressed in a variety of cells and tissues such as placenta, brain, testis, 
      liver, kidney, skeletal muscle, primary blood mononuclear cells and cancer cells. 
      We observed previously that the enhancer region of the HP2K gene, which has been 
      identified in the 5'-flanking region between -1265 and -1329, could respond to 
      serum stimulation following the transfection of human choriocarcinoma BeWo cells 
      with HP2K promoter-luciferase constructs. The HP2K enhancer region also contains 
      two copies of the hypoxia-inducible factor-1 (HIF-1) binding motif (5'-ACGTG-3'). 
      In this study we performed characterization of the HP2K gene expression in 
      response to hypoxic conditions. Both electrophoretic mobility shift and 
      co-transfection assays of the HP2K promoter-luciferase reporter with HIF-1 
      expression vectors indicated that HIF-1 binds to the hypoxia-responsive element 
      (HRE) of HP2K, thereby upregulating its gene expression. In addition, we 
      demonstrated using site-directed mutagenesis that a complete tandem repeat of the 
      HIF-1 binding motif with a 4-bp interruption is required for full induction of 
      HP2K expression (up to 22-fold) under hypoxic conditions, and that this response 
      is much stronger than that of the erythropoietin (EPO) gene. These results 
      suggest that the sequence 5'-ACGTGNNNNACGTG-3' in the HP2K enhancer is the 
      authentic HRE consensus motif that mediates increased transcription, under 
      hypoxic conditions, via HIF-1.
FAU - Fukasawa, Masashi
AU  - Fukasawa M
AD  - Department of Microbiology, National Defense Medical College, 3-2 Namiki, 
      Tokorozawa, Saitama 359-8513, Japan.
FAU - Tsuchiya, Terumasa
AU  - Tsuchiya T
FAU - Takayama, Eiji
AU  - Takayama E
FAU - Shinomiya, Nariyoshi
AU  - Shinomiya N
FAU - Uyeda, Kosaku
AU  - Uyeda K
FAU - Sakakibara, Ryuzo
AU  - Sakakibara R
FAU - Seki, Shuhji
AU  - Seki S
LA  - eng
PT  - Journal Article
PL  - England
TA  - J Biochem
JT  - Journal of biochemistry
JID - 0376600
RN  - 0 (HIF1A protein, human)
RN  - 0 (Hypoxia-Inducible Factor 1, alpha Subunit)
RN  - 0 (Oligonucleotides)
RN  - 0 (Protein Isoforms)
RN  - 0 (Transcription Factors)
RN  - 11096-26-7 (Erythropoietin)
RN  - EC 1.13.12.- (Luciferases)
RN  - EC 2.7.1.105 (Phosphofructokinase-2)
SB  - IM
MH  - Amino Acid Motifs
MH  - Base Sequence
MH  - Cell Line
MH  - Cell Line, Tumor
MH  - Cell Nucleus/metabolism
MH  - Enhancer Elements, Genetic
MH  - Erythropoietin/genetics/metabolism
MH  - Gene Expression Regulation
MH  - Genetic Vectors
MH  - Glycolysis
MH  - Humans
MH  - *Hypoxia
MH  - Hypoxia-Inducible Factor 1, alpha Subunit
MH  - Luciferases/metabolism
MH  - Molecular Sequence Data
MH  - Oligonucleotides/chemistry
MH  - Phosphofructokinase-2/*chemistry/physiology
MH  - Placenta/*enzymology
MH  - Promoter Regions, Genetic
MH  - Protein Isoforms
MH  - Protein Structure, Tertiary
MH  - Time Factors
MH  - Tissue Distribution
MH  - Transcription Factors/*genetics/metabolism
MH  - Transcription, Genetic
MH  - Transfection
MH  - Up-Regulation
EDAT- 2004/12/16 09:00
MHDA- 2005/07/19 09:00
CRDT- 2004/12/16 09:00
PHST- 2004/12/16 09:00 [pubmed]
PHST- 2005/07/19 09:00 [medline]
PHST- 2004/12/16 09:00 [entrez]
AID - 136/3/273 [pii]
AID - 10.1093/jb/mvh137 [doi]
PST - ppublish
SO  - J Biochem. 2004 Sep;136(3):273-7. doi: 10.1093/jb/mvh137.